Staining solution for cell staining and preparation method thereof
A staining solution and cell technology, applied in the field of biochemical detection, can solve problems such as poor cell effect
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Embodiment 1
[0210] Embodiment 1 determines the effective color concentration C of AO AO1 Range and AO channel effective color rendering exposure time T AO1 scope
[0211] 1.1 Obtain live cell samples of MCF-7 stained by AO
[0212] Freshly digested MCF-7 cells (purchased from ATCC) were collected and resuspended in serum-containing cell culture medium to obtain live MCF-7 cell samples (cell viability >95%).
[0213] Different concentrations of AO staining solution were prepared by using staining reagent AO (Beijing Suo Laibao, product number A8120) and PBS buffer, and the AO staining solution was adjusted to pH 7.5. Among them, the AO concentration of each AO staining solution was 0.04 μg / mL, 0.2 μg / mL, 1.0 μg / mL, 5.0 μg / mL, and 25.0 μg / mL, respectively.
[0214] Five groups of MCF-7 living cell samples were taken, numbered sequentially according to the group, and each group had 5 parallel samples, and the volume of each sample was 20 μL. According to the corresponding relationship sh...
Embodiment 2
[0225] Example 2 screens out the preferred range of AO effective color development concentration and the preferred range of effective color development exposure time of AO channel
[0226] 2.1 Obtain live cell samples of MCF-7 stained by AO
[0227] Obtain MCF-7 living cell samples, the method refers to step 1.1 of Example 1.
[0228] Different concentrations of AO staining solution were prepared, and the AO staining solution was adjusted to pH7.5. Wherein, the AO concentration of the AO staining liquid is at the effective color development concentration C of the dyeing reagent AO initially judged in Example 1. ao1 Selected within the value range of 0.3μg / mL, 1.2μg / mL, 4μg / mL respectively.
[0229] Three groups of MCF-7 living cell samples were taken, numbered in sequence according to the group, and three parallel samples were set up for each group, and the volume of each sample was 20 μL. According to the corresponding relationship shown in Table 3, 20 μL of AO staining so...
Embodiment 3
[0238] Embodiment 3 determines the effective color concentration C of PI PI1 Range and PI channel effective color rendering exposure time T PI1 scope
[0239]3.1 Obtain PI-stained MCF-7 dead cell samples
[0240] Freshly digested MCF-7 cells were collected, added with a final concentration of 0.1% Triton, shaken and mixed for 1 minute, and dead MCF-7 cells (cell viability rate 0%) were obtained.
[0241] Different concentrations of PI staining solution were prepared by using staining reagent PI (Beijing Suo Lai Bao, product number C0080) and PBS buffer solution, and adjusted to pH 7.5. Wherein, the PI concentrations of each PI staining solution were 10 μg / mL, 50 μg / mL, 250 μg / mL, 1000 μg / mL, and 5000 μg / mL, respectively.
[0242] Five groups of MCF-7 dead cell samples were taken and numbered sequentially by group, with 5 parallel samples in each group, and the volume of each sample was 20 μL. According to the corresponding relationship shown in Table 5, add 20 μL of PI sta...
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